1.Repairing critical-size segmental defects:morphology of tissue engineering bone scaffolds and its effects on cell loading
Chinese Journal of Tissue Engineering Research 2014;(29):4717-4723
BACKGROUND:Tissue engineering bone application for repairing critical-size segmental defects is stil in research stage. The ideal construction methods have not yet been found.
OBJECTIVE:To review the research literatures on tissue engineering bone scaffold material, its shape and effect on the loading of seeding cells, seek appropriate engineered bone scaffolds which are capable of loading a large number of cells effectively and probably, and provide a new way of repairing segmental bone defects.
METHODS:The first author performed a data retrieval of PubMed and Wanfang databases from 1994 to 2013, to search the articles addressing the construction method of tissue engineering bone scaffold, and reviewed the literatures systematical y.
RESULTS AND CONCLUSION:A total of 379 references were retrieved, including 161 articles in Chinese and 218 articles in English. According to the inclusion and exclusion criteria, 53 articles were final y involved in the analysis. The analysis results indicated that, the needed volume of bone tissue engineering scaffolds for critical-sized section bone defect reconstruction is big, which needs to load a huge number of seed cells. If there is no suitable forms and shapes for celladhesion, the property of so-cal ed engineered bone is similar to pure artificial bone implants. The effective load of seed cells on engineering bone scaffold material and keeping the activity is the first step in clinical practice, as wel as the important guarantee for loading bioactive seed cells. Hence, a more simple and accurate detection method for loading cellquantity is needed. Looking into the retrieved content, effective load cellquantity and its bioactivity are detected by indirect methods, supporting the effectiveness of cellseeding. Some methods can guarantee the cellquantity and seeding pattern, the real load is unknown as wel as the activity. Fabricating engineering bone scaffold into special form and shape are easy to effective seeding, proliferation and maintaining the biomechanical performance, inducing osteogenesis, and final y detecting the load cellquantity and activity on the scaffold through the simple and direct method.
2.CONSTRUCTION AND EXPRESSION OF HUMAN Axud1 GENE WITH HA EPITOPE TAG IN LUNG ADENOCARCINOMA SPC-A1 CELL LINE
Fan DENG ; Zhipeng ZOU ; Chunlu YANG
Medical Journal of Chinese People's Liberation Army 2001;0(07):-
To construct the eukaryotic vector that expresses the fusion protein of Axud1 and influenza virus hemagglutin HA epitope tag, the total RNA was isolated from the peripheral blood lymphocytes, and reverse transcription reaction was used to amplify the full length of human Axud1 cDNA. PCR product of Axud1 was then amplified using specific primers containing HA epitope sequence, and inserted into eukaryotic expression plasmid pcDNA3.1(+)digested with BamH Ⅰand Xba Ⅰ. The recombinant plasmid was identified by polymerase chain reaction, restriction endonuclease mapping and sequencing, and then transfected into human lung adenocarcinoma SPC-A1 cell lines.The fusion HA-Axud1 protein expression in anti-G418 clones was verified by Western blot. This study might be instrumental in further study of the function of Axud1 protein in tumor cells.
3.Suppression of ABCE1 Leads to Decreased Cell Proliferation and Increased Apoptosis in 95-D/NCI-H446 Lung Carcinoma Cells
Maogen ZHENG ; Ying GAO ; Bo HUANG ; Dali TIAN ; Chunlu YANG
Progress in Biochemistry and Biophysics 2009;36(11):1475-1482
ATP-binding cassette protein E (ABCE1) has been annotated as an Rnase L inhibitor in eukaryotes. Previous study showed that the overexpression of ABCE1 was related with the occurrence and clinical stage of lung adenocarcinoma. As an initial investigation into the novel functions of ABCE1, siRNA-expressing vectors targeting sites of the ABCE1 gene were constructed from RNAi-Ready pSIREN-DNR-DsRed-Express vector. Cultured 95-D and NCI-H446 lung carcinoma cells were transfected with the siRNA-expressing vectors using FuGENE 6 and transfection efficiency was determined by using fluorescence microscopy. The expression level of ABCE1 protein was determined by Western blot and immunofluorescence staining. Cell viability was determined by MTT, cell cycle was analysed by flow cytometry.The apoptotic rate was observed by ELISA. Fluorescence microscopy showed a satisfactory transfection efficiency which was about 42.70%. Cell viability and the growth fraction were markedly suppressed, whereas the apoptosis was significantly increased in SiRNA-95-D and SiRNA-NCI-H446 cells than controls(P< 0.05). It can be concluded that the siRNA targeting ABCE1 gene shows a dramatic inhibitory effect on RNA transcription and protein expression and a promoting effect on the apoptosis in 95-D/NCI-H446 cells, which offers a reliable base for the further in vivo experiment.
4.A study on the effect of great saphenous vein interruption on distally saphenous neurocutaneous pedicled skin flap
Peng JI ; Chunlu YANG ; Weixiong HUANG ; Xinlong ZOU
Chinese Journal of Postgraduates of Medicine 2016;(1):54-56
Objective To explore the effect of great saphenous vein interruption on distally saphenous neurocutaneous pedicled skin flap. Methods Fifty-two patients with skin and soft tissue lost on ankle received neoplasty using distally crural saphenous neurocutaneous pedicled skin flap. The patients were divided into two groups: the patients in interruption group (25 patients) were treated with great saphenous vein interruption on distally saphenous neurocutaneous pedicled skin flap, the patients in conventional group (27 cases) were treated without saphenous vein interruption. Results Primary healing: 15 patients (55.56%, 15/27) in conventional group, 21 patients (84.00%,21/25) in interruption group. With effusion: 17 patients (62.96%,17/27) in conventional group, 7 patients(28.00%,7/25) in interruption group. With venous crisis: 10 patients (37.04%,10/27) in conventional group, 2 patients (8.00%,2/25) in interruption group. There was statistical significance between two groups on the above 3 indexes (P <0.05). With infection: 7 patients (25.93%,7/27) in conventional group; 4 patients(16.00%,4/25) in interruption group. There was no statistical significance between two groups (P>0.05). Conclusions Great saphenous vein interruption could relieve swelling, reduce effusion and have higher primary healing rate in neoplasty using distally crural saphenous neurocutaneous pedicled skin flap compared with the conventional method, which greatly reduce the pain and medical expenses of the patients.
5.Establishing an animal model of delayed onset muscle soreness and its histomorphologic observation
Yuan WEI ; Chunlu FANG ; Liangming LI ; Wenhua XING ; Zeyi YANG
Chinese Journal of Tissue Engineering Research 2015;(29):4645-4651
BACKGROUND:Delayed onset muscle soreness is closely related to skeletal muscle micro-injury, but the exact mechanism of skeletal muscle micro-injury is not yet clear. OBJECTIVE:To observe the histomorphological and ultrastructure changes of skeletal muscle micro-injury models induced by eccentric exercise. METHODS: Forty male Sprague-Dawley rats were randomly divided into quiet control group, immediately after exercise group, post-exercise 24 hours group, post-exercise 48 hours group and post-exercise 72 hours group. In the latter four groups, the rats were subjected to intermittent running on the-16° slope at a speed of 16 m/min: 5 minutes movement, 2 minutes rest and totaly 120 minutes. Rats in the latter four groups were observed immediately, at 24, 48, 72 hours after exercise. RESULTS AND CONCLUSION:After eccentric exercise, the morphology and ultrastructure of rat’s skeletal muscle were both changed to different extents, and Desmin and Vimentin were dyed off for anti-desmin antibody staining at varying degrees. It indicates that one-time eccentric exercise can cause delayed skeletal muscle micro-injury.
6.Recent advance in surgical treatment of pancreatic neuroendocrine tumors
Min YANG ; Chunlu TAN ; Nengwen KE ; Xubao LIU
Chinese Journal of Endocrine Surgery 2017;11(3):241-244
Pancreatic neuroendocrine tumors (P-NETs) are a group of heterogeneous tumors,including functional and nonfunctional ones.With the enhancement of clinicians' awareness about this disease and the improvement of imaging diagnostic techniques,the incidence of P-NETs has obviously increased in the past years.Based on the mitotic counting and Ki-67 positive index,the grading classification is of great value for the diagnosis,treatment and even prognosis of P-NETs.P-NETs are a group of malignant tumors with inert biological behaviors,whose surgical resection rate and long-term survival is much better than those of pancreatic ductal adenocarcinoma.P-NETs have different malignant potentials.Clinicians need to develop a comprehensive treatment plan in combination with the patient's symptoms,tumor grading classification and TNM staging information.Surgery is the only curable way to cure P-NETs.Even if radical resection is not suitable,palliative surgery may alleviate the patients,symptoms,and even prolong their survival time.According to the tumor location,size,quantity,degree of grading,local invasion and distant metastasis,different surgical procedures should be selected.
7.Analysis of Clinical Features and Risk Factors of Recurrence after Choledocholithotomy
Song HU ; Shuyan FANG ; Guangfu JIN ; Han LI ; Guang YANG ; Hui XIA ; Chunlu MU
Progress in Modern Biomedicine 2017;17(22):4299-4302
Objective:To explore the clinical features of recurrence after choledocholithotomy and to analyze the risk factors.Methods:The clinical data of 730 patients with choledocholithiasis who were treated in our hospital from January 2005 to July 2016 were analyzed retrospectively,550 cases who were received choledocholithotomy were defined as laparotomy group,30 cases with laparoscopic common bile duct exploration (LCBDE) were defined as the LCBDE group,and 150 cases with endoscopic sphincterotomy (EST) were defined as EST group.The recurrence rate of the three groups were compared.The patients of three groups were divided into recurrence group (n=227) and non recurrence group (n=503) according to the recurrent situation,then the clinical features and risk factors of recurrent patients were analyzed by univariate and multivariate Logistic regression analysis.Results:The recurrence rate of EST group was 38.67%,which was significantly higher than that of LCBDE group with 26.67% and the laparotomy group with 29.27%,and there was statistical difference (P<0.05).The results of univariate analysis showed that there were statistically significant differences in age,history of HBV infection,jaundice,abnormal total bilirubin,peripapillary diverticulum,biliary infection,biliary stricture,papillary stenosis,sphincter of Oddis dysfunction,history of biliary surgery,cholecystectomy,bile duct diameter ≥ 15 mm,bile duct angle ≤120°,operation type,stone quantity ≥ 2 grains,stone diameter ≥ 10 mm,with or without gallstones (P<0.05).The results of Logistic multivariate regression analysis showed that age,having peripapillary diverticulum,having history of biliary surgery,bile duct diameter ≥ 15 mm,stone quantity ≥ 2grains and EST operation type were the independent risk factors of the recurrence after choledocholithotomy (P<0.05).Conclusion:There are many risk factors of recurrence after choledocholithotomy,and operation method should be based on the size and the number of the stones,and the constitution of patients.Preventive measures should be strengthened to control the recurrence after choledocholithotomy.
9.Early orogastric feeding compared with nasoenteric tube enteral nutrition in postoperative pancreaticoduodectomy patients
Wei YANG ; Chunlu TAN ; Kezhou LI ; Yan YANG
Chinese Journal of General Surgery 2017;32(9):738-741
Objective To study the postoperative nutritional effects of early orogastric feeding compared with nasoenteric tube enteral nutrition within postoperative 24-48 h in pancreaticoduodenectomy (PD) patients.Methods The clinical data of 87 PD patients from June 2013 to January 2015 in Huaxi Hospital was analyzed retrospectively.Patients were divided into orogastric enteral nutrition group and nasoenteric tube enteral nutrition group.Patients' tolerance,hospital stay,mortality,and major complications associated with PD were compared.Results The average time of bowel function recovery (P <0.001) and average hospital stay (P=0.017) in the nasoenteric tube enteral nutrition group were significantly shorter than those in the orogastric enteral nutrition group.The differences of patients' tolerance were not statistically different (P =0.089).The differences of major complications and mortality were not statistically different (P =0.745,P =1.00).Conclusions Nasoenteric tube enteral nutrition in after PD patients improves nitrogen balance,promotes bowel function recovery and makes hospital stay shorter.
10.Effects of corrugated tissue engineered bone scaffold on cell seeding and osteogenesis
Xianli ZENG ; Chunlu YANG ; Jiang LI ; Yuan XU ; Shuo WANG ; Peng WU ; Cuifang WANG ; Yanli DING ; Xinli ZHENG
Chinese Journal of Tissue Engineering Research 2014;(25):3953-3960
BACKGROUND:The effects of engineered bone scaffold containing seeding cels with different shapes to repair bone defect are varied, while the loaded cellquantity is the important factor influencing the curative effect, but which is rarely reported. OBJECTIVE:By preparing self-made corrugated tissue-engineered bone scaffold and other three forms of bone tissue engineering scaffolds, to study the quantity of loaded cels on different scaffolds and osteogenesis of corrugated tissue-engineered bone scaffold so as to discuss the advantages and features of self-made corrugated tissue-engineered bone scaffold. METHODS: (1) Experimentin vitro: There were four kinds of scaffolds with the same volume and samples: calcium phosphate cement (CPC) corrugated surface scaffold group, smooth surface scaffold group, cylindrical scaffold group and porous cylindrical scaffold with holow tubes group, in which the latter three groups are control ones. A certain volume with same density of rabbit bone marrow mesenchymal stem cels (BMSCs) suspension after osteogenesis induction was seeded onto the scaffolds. After incubation, culture, digestion and colection, cellquantity was counted, absorbance value was finaly detected and cellactivity was proofed by alkaline phosphatase and alizarin red staining. (2) Experimentin vivo: New Zealand rabbits were randomly and equaly divided into recombinant human bone morphogenetic protein-2 (rhBMP-2)/CPC/BMSCs corrugated scaffold group, pure CPC corrugated scaffold group and cancelous bone implant group. Three kinds of scaffold implants with the same volume were inserted into the area between rabbit’s L5, 6 transverse processes bilateraly. At 4, 8, 12 weeks postoperatively, gross and histological observation was performed. RESULTS AND CONCLUSION:(1)Experimentin vitro: The drip of cellsuspension steadily stayed on the surface of corrugated scaffold because of corrugated shape groove and the surface tension of the liquid. The amount of cels per sample digested down from the CPC corrugated surface scaffold was significantly more than that from the other three groups (P < 0.05), while the absorbance values did not differ (P > 0.05). (2) Experimentin vivo: At each time point the osteogenesis quantity of rhBMP-2/CPC/BMSCs corrugated scaffold group was more than that of the pure CPC corrugated scaffold group (P < 0.05), while there was no difference from the cancelous bone implant group (P > 0.05). These findings indicate that the characteristics of the self-made corrugated engineered bone scaffold are beneficial to seed cellloading, which supports a large number of osteogenesis and provides feasibility to promote the healing of segmental bone defects.